Method of real time detecting No.21 human chromosome number by quantitative PCR technology

A chromosome number, chromosome technology, applied in the field of molecular biology

Inactive Publication Date: 2005-11-09
ATTACHED OBSTETRICS & GYNECOLOGY OSPITAL MEDICALCOLLEGE ZHEJIANG UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0007] So far, there have been no reports at home and abroad on the use of multiplex real

Method used

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Experimental program
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Embodiment 1

[0030] 1. Chromosome molecular marker selection

[0031] The target sequence on chromosome 21 is:

[0032] GTTGATGGGC TTGCGCTTA TCTGCTTGTCC CTTCTCCCAT CTGATTTCTT GTGGAGCCCA TGG

[0033] Among them, the upstream primer sequence: 5'-GTTGATGGGCTTGCGCTTAT-3'

[0034] Downstream primer sequence: 5'-CCATGGGCTCCACAAGAAAT-3'

[0035]Taqman probe sequence: 5'-Fam-TGCTTGTCCCTTCTCCCATCT-Tamra-3'

[0036] The internal reference sequence on chromosome 1 is:

[0037] TCAACATGGG CCCCACATGG TCCTAGTTTC CCCGGCTCAT TATGCTTCAG GCACACTGGCTTTCTTGC

[0038] Among them, the upstream primer sequence: 5'-TCAACATGGGCCCCACAT-3'

[0039] Downstream primer sequence: 5'-GCCAGTGTGCCTGAAGCA-3'

[0040] Taqman probe sequence: 5'-Vic-TCCTAGTTTTCCCCGGCTCATT-Tamra-3'

[0041] 2. Specimen Preparation

[0042] Specimens were 0.2ml of fresh EDTA anticoagulated whole blood or 10ml of amniotic fluid (centrifuged at 1000rpm for 15min to pellet fetal cells and diluted in 0.2ml of PBS buffer). D...

Embodiment 2

[0049] normal blood sample

[0050] According to the detection range determined by us, the detection sensitivity of the method of the present invention to trisomy 21 is 100%, the specificity is 95.6%, the false positive rate is 4.4%, and the false negative rate is 0.

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Abstract

A method for detecting the number of human chromosomes NO.21 by real-time quantitative PCR technique in order to diagnoise the triple-21 syndrome includes such steps as using the genome-specific single-cope sequence as the molecular marker of chromosome, choosing part of non-polymorphic fragments of DSCR4 gene as target molecular marker, choosing part of fragments of RABIF gene as internal reference molecular marker, designing two pairs of primers and relative Taqman probes, amplifying said two sequences in a single tube by PCR, determining the relative value deltact, and determining the number of chromosomes No.21.

Description

technical field [0001] The invention belongs to molecular biology, and relates to a method for detecting the number of chromosomes, in particular to a method for detecting the number of human chromosome 21 by using multiple real-time quantitative PCR technology. Background technique [0002] Trisomy 21 (referred to as trisomy 21, also known as Down syndrome or congenital stupidity) is the most common chromosomal disease in humans, with an incidence of about 1 / 600 live births[1], manifested as 21 The number of chromosomes is abnormal, that is, the patient has one more than the normal two (or part of the translocation chromosome 21). Patients with this disease have severe mental retardation, and are prone to heart disease, leukemia, low immune function, and high early mortality in children. At present, there is no effective treatment for the disease, and prenatal diagnosis is mainly used to avoid the birth of defective children. The classic prenatal diagnosis of congenital s...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 吕时铭朱宇宁裘俭朱波尤建飞
Owner ATTACHED OBSTETRICS & GYNECOLOGY OSPITAL MEDICALCOLLEGE ZHEJIANG UNIV
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